WO2013071301A1 - Test diagnostique à base de papier - Google Patents

Test diagnostique à base de papier Download PDF

Info

Publication number
WO2013071301A1
WO2013071301A1 PCT/US2012/064856 US2012064856W WO2013071301A1 WO 2013071301 A1 WO2013071301 A1 WO 2013071301A1 US 2012064856 W US2012064856 W US 2012064856W WO 2013071301 A1 WO2013071301 A1 WO 2013071301A1
Authority
WO
WIPO (PCT)
Prior art keywords
substrate
zone
diagnostic device
blood
agglutination
Prior art date
Application number
PCT/US2012/064856
Other languages
English (en)
Inventor
Sergy S. SHEVKOPLYAS
Xiaoxi YANG
Julie Kanter WASHKO
Nathaniel Zane PIETY
Original Assignee
The Administrators Of The Tulane Educational Fund
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Administrators Of The Tulane Educational Fund filed Critical The Administrators Of The Tulane Educational Fund
Priority to US14/357,074 priority Critical patent/US20140295472A1/en
Priority to KR1020147015765A priority patent/KR101659529B1/ko
Priority to EP12848168.6A priority patent/EP2776831A4/fr
Priority to CA2855363A priority patent/CA2855363C/fr
Priority to CN201280062780.1A priority patent/CN104126120B/zh
Publication of WO2013071301A1 publication Critical patent/WO2013071301A1/fr
Priority to US15/215,107 priority patent/US10908152B2/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G01N33/521—Single-layer analytical elements
    • G01N33/523—Single-layer analytical elements the element being adapted for a specific analyte
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G01N33/525—Multi-layer analytical elements
    • G01N33/526—Multi-layer analytical elements the element being adapted for a specific analyte
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
    • G01N30/92—Construction of the plate
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/49—Blood
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/66—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood sugars, e.g. galactose
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/795—Porphyrin- or corrin-ring-containing peptides
    • G01N2333/805—Haemoglobins; Myoglobins

Definitions

  • the present invention relates to simple, low-cost, rapid paper-based diagnostic devices and their methods of use.
  • Microfluidic paper-based devices are typically small, portable and easily fabricated from inexpensive materials and delivered to remote, resource-limited locations.
  • ⁇ may be easily fabricated by printing patterns onto paper with a solid ink (wax) printer and melting the ink to create hydrophobic barriers spanning through the entire thickness of the paper substrate.
  • the ⁇ use the paper as a fluidic substrate, and utilize the wicking/capillary properties of the paper to transport the biological sample from a sample deposit region.
  • These devices do not typically require complex laboratory equipment, and thus are well-suited for diagnostic applications in clinical practice generally, and particularly in developing countries, in emergency situations and home health care situations.
  • colorimetric assays Many of these ⁇ run colorimetric assays.
  • the use of colorimetric assays for analysis of biological fluids is generally attractive because these assays produce a visual readout and are usually simple to perform, stable, and inexpensive.
  • the biological sample reacts with reagents deposited within a test readout zone, and the reaction produces a detectable color.
  • traditional colorimetric assays are limited to optically transparent samples (e.g., water, urine, pre- separated blood plasma). If a non-transparent sample is used, then the color of the sample can interfere with the detection of the developed color.
  • Blood plasma is commonly used as the biological sample because its composition is exceptionally informative about the pathological processes affecting organs and tissues throughout the body. For example, the detection of non-esterified fatty acids, glucose, heparin and lysophosphatic acid are performed by testing blood plasma.
  • the plasma in order to use blood plasma in the colorimetric assays, it is beneficial for the plasma to be first separated from the whole blood. Blood plasma separation is a particularly important step for a colorimetric assay because the intense color of the red blood cells (“RBCs") in the whole blood may interfere with quantification of the results of the diagnostic colorimetric assays.
  • RBCs red blood cells
  • Including the plasma separation step into the design of colorimetric ⁇ would transform them into fully integrated diagnostic devices and thus significantly increase their versatility by eliminating the need for a separate sample preparation step which often requires expensive, bulky equipment and specially trained personnel. These fully integrated ⁇ would be able to analyze samples of whole blood taken directly in the field and simply placed on the agglutination zone of the device. Integrated plasma separation could make colorimetric ⁇ suitable for many more applications and situations in which one may use colorimetric methods to test the multitude of clinically relevant biomolecules present in human blood plasma, while controlling for the interference from the deep-red color presented by the RBCs. Thus, there is a need for an innovation in ⁇ in order to allow for point-of-case diagnostics with the ability for automated quantification.
  • ⁇ may also be used to detect the presence of sickling hemoglobin in a blood sample (e.g., to diagnose sickle cell disease).
  • Hemoglobin (Hb) is the iron-containing oxygen-transport protein in RBCs. Each molecule of hemoglobin consists of four globin chains: fetal hemoglobin (Hb F) has two and two y chains, and adult hemoglobin (Hb A) has two a and two ⁇ chains.
  • Mutations of the genes controlling the globin chain production include structural variants that change the amino acid sequence and produce aberrant forms of Hb, and mutations that lower or eliminate production of globin chains (thalassaemias).
  • deoxy-Hb S changes conformation such that the hydrophobic patch at the site of the valine replacement on a ⁇ chain of one Hb S molecule binds to a complementary hydrophobic site on & ⁇ chain of another Hb S molecule.
  • the polymerization of Hb S in an anaerobic environment gives RBCs a distorted, sickled shape.
  • Those who inherit only one copy of Hb S and possess the other copy of the gene encoding for the normal Hb A carry the sickle cell trait (SCT), but are generally considered healthy, although with a higher risk for venous thromboembolism and renal medullary carcinoma.
  • SCT sickle cell trait
  • Hb S Those w r ho inherit two copies of Hb S (genotype Hb SS) develop sickle cell anemia, the most prevalent form of sickle cell disease (SCD).
  • SCD sickle cell disease
  • Newborn screening has been the single greatest advance in the treatment of SCD in high- income countries.
  • SCD is diagnosed primarily through hemoglobin electrophoresis (HE), but also using high performance liquid chromatography (HPLC) and isoelectric focusing (IEF) testing, which exploit the differences in the electric charge of Hb variants to detect their presence in RBCs of the patient.
  • HPLC high performance liquid chromatography
  • IEF isoelectric focusing
  • An aspect of the present invention includes a diagnostic device comprising a substrate having pores, an agglutination zone, and a test readout zone, wherein said agglutination zone is functionalized with an agglutinating agent.
  • a further aspect of the present invention includes a method of diagnosing a disease or condition comprising the steps of providing a diagnostic device comprising a substrate having pores, an agglutination zone, a test readout zone, and wherein said agglutination zone is functionalized with an agglutinating agent and said test readout zone is functionalized with an assay reagent; depositing a blood sample onto said agglutination zone; allowing said blood sample to develop; and observing said test readout zone.
  • a still further aspect of the present invention includes a method of diagnosing a disease or condition comprising the steps of providing a diagnostic device comprising a substrate having pores and an agglutination zone; mixing a volume of blood sample with a volume of agglutinating agent; depositing a droplet of said mixture onto said agglutination zone; allowing said droplet to develop and create a blood stain pattern on said substrate; and observing said blood stain pattern.
  • a still further aspect of the present invention includes a method of diagnosing a disease or condition comprising the steps of: providing a diagnostic device comprising a substrate having pores, wherein said substrate further comprises an agglutination zone and a test readout zone, and wherein said agglutination zone is functionalized with an agglutinating agent and said test readout zone is functionalized with an assay reagent; depositing a blood sample onto said agglutination zone; allowing said blood sample to develop; and observing said test readout zone.
  • a still further aspect of the present invention includes a method of diagnosing a disease or condition comprising the steps of: providing a diagnostic device comprising a substrate having pores, wherein said substrate further comprises an agglutination zone; mixing a volume of blood sample with a volume of agglutinating agent; depositing said droplet onto said agglutination zone; allowing said droplet to develop and create a blood stain pattern on said substrate; and observing said blood stain pattern.
  • a still further aspect of the present invention includes a system for diagnosing a disease or condition comprising: a substrate having pores, wherein said substrate further comprises: an agglutination zone and a test readout zone; wherein said agglutination zone is functionalized with an agglutinating agent; an optical image capture device capable of capturing an image of said test readout zone; and computer software capable of analyzing said image.
  • a still further aspect of the present invention includes a system for diagnosising a disease or condition comprising: a substrate having pores, wherein said substrate further comprises an agglutination zone; and a sample deposited on said agglutination zone, wherein said sample is comprised of a mixture of whole blood and an agglutinating agent; an optical image capture device capable of capturing an image of said substrate; and computer software capable of analyzing said image.
  • a still further aspect of the present invention includes a device for diagnosing a disease or condition comprising: a means for receiving a blood sample; a means for agglutinating red blood cells of said blood sample; a means for transporting plasma of said blood sample away from said receiving means; and a means for determining the presence of an analyte in said plasma.
  • a still further aspect of the present invention includes a device for diagnosing a disease or condition comprising: a means for receiving a sample comprised of: whole blood mixed with a means for agglutinating said whole blood; a means for transporting soluble forms of Hb of said sample away from said receiving means and creating a blood stain pattern; a means for scanning said blood stain pattern; and a means for correlating said scanned blood stain pattern with said diagnosis of said disease or condition.
  • FIG. 1A illustrates the known RBC characteristic of deformability when RBCs are attempted to be separated from whole blood.
  • FIG. IB illustrates the known method of filtering plasma from RBCs in whole blood.
  • FIG. 1C illustrates the use ofRBC agglutination to filterplasma from RBCs in whole blood.
  • FIG. 2A illustrates the result of spotting whole blood on untreated paper.
  • FIG. 2B illustrates the result of spotting whole blood on paper treated with agglutinating antibodies anti-A,B.
  • FIG. 2C illustrates the relation between volume of the whole blood sample and the radius of the whole blood spot, radius of the agglutinated RBC spot and the width of the plasma band.
  • FIG. 3 illustrates the design of a ⁇ with integrated blood plasma separation.
  • FIG. 4A illustrates the operation of a ⁇ with integrated blood plasma separation.
  • FIG. 4B illustrates a ⁇ ) with integrated blood plasma separation after the reaction at the test zones has occurred.
  • FIG. 5 illustrates the quantification of glucose concentration in whole blood samples using a ⁇ with integrated blood plasma separation.
  • FIG. 6 illustrates a schematic diagram of the use of a paper-based hemoglobin solubility assay utilizing agglutination.
  • FIG. 7A illustrates the blood stain patterns created during the use of a paper-based hemoglobin solubility assay utilizing agglutination.
  • FIG. 7B illustrates red color intensity profiles quantifying the blood stain patterns shown in FIG. 7A.
  • FIG. 8 A illustrates color intensity profiles of normal (Hb AA), SCT (Hb AS) and SCD (Hb SS, Hb Sf, or Hb SC) blood samples.
  • FIG. 8B illustrates normalized color intensity values at 5mm for each sample of FIG. 8A.
  • One aspect of the present invention provides a diagnostic device and its method of use for separating blood plasma from red blood cells (RBCs) in small samples of whole blood contained entirely within a ⁇ .
  • RBCs red blood cells
  • normal healthy RBCs 100 are extremely deformable, and can easily pass through a substrate 101 with pores 102 smaller than the diameter of the RBCs 100.
  • the substrate 100 may be paper, specifically chromatography paper, cloth, string or any other material with wicking or capillary properties.
  • the diameter of the smallest pore 102 that that a RBC 100 could pass through depends on the volume and surface area of the cell, but is approximately 2.5 ⁇ for a normal human RBC. Therefore, a substrate 101 with pores 102, each with a diameter (d) of less than 2.5 ⁇ , provides a fairly straightforward means for separating plasma from RBCs 100 in the whole blood samples, as illustrated in Figure IB.
  • the present invention utilizes BC agglutination to increase the effective size of RBCs by forming large multi-cellular agglutinated RBCs 103 that can be filtered out using a substrate 101 with pores 102 significantly larger than 2.5 ⁇ , and thus produce purified plasma at a much higher volumetric flow rate.
  • Agglutination generally, is the clumping of particles, such as RBCs, to create a larger particle. Agglutination can be caused by the addition of an agglutinating agent, or alternatively by a change in temperature.
  • RBCs Because of agglutination, RBCs form agglutinated RBCs 103 that are too large to pass through the pores 102 within the substrate 101 of the ⁇ AD. As a result, agglutinated RBCs 103 become entangled in the substrate 101 and thus separate from the plasma in a whole blood sample. The plasma, which passes through the pores 102 of the substrate 101 , is then wicked outwardly through the substrate 101. In the present invention, the pores 102 in the substrate 101 are sufficiently small to efficiently filter out the agglutinated RBCs 103, yet large enough to enable adequately high rates of blood plasma flow for completing a colorimetric assay.
  • Agglutination can be initiated by adding an agglutinating agent, such as agglutinating antibodies (anti- ⁇ , ⁇ ) to whole blood.
  • agglutinating antibodies anti- ⁇ , ⁇ are monoclonal antibodies of the immunoglobulin class IgM, which selectively bind to antigen A and antigen B present on the surface of human RBCs.
  • Direct agglutination of RBCs by anti- ⁇ , ⁇ antibodies occurs when either A or B, or both A and B antigens are present on the surface of RBCs (blood types A, B and AB).
  • FIG. 2A depicts the visual appearance of a whole blood sample spotted onto paper substrate treated with phosphate buffered saline (for control) and FIG. 2B depicts the visual appearance of a whole blood sample spotted onto paper treated with agglutinating antibodies. Comparing FIG. 2Ato FIG. 2B, one can see that the whole blood sample 201 spotted on paper pre-treated with phosphate buffered saline (for control) behaved as a uniform phase with RBCs and plasma wicking through the paper without separation.
  • the plasma separated from agglutinated RBCs 202 (that became entangled in the paper fibers), creating a plasma band 203 around the agglutinated RBCs 202.
  • the plasma band 203 spread significantly further than agglutinated RBCs 202 or the whole blood sample 201 on phosphate buffered saline treated paper.
  • FIG. 3 illustrates the design of a ⁇ 300 integrating blood plasma separation from whole blood using agglutination.
  • the pattern of the ⁇ 300 includes the agglutination zone 301 in the center region and four test readout zones 302, 303, 304, 305 on the periphery of the ⁇ 300.
  • the aforementioned data (illustrated in FIG. 2C) was used to determine the optimal shape and size of the ⁇ 300 for effectively retaining agglutinated RBCs within the central part of the ⁇ 300 and enabling the flow of a sufficient amount of separated plasma into the test readout zones 302, 303, 304, 305 on the periphery.
  • the optimal distance between the center of the agglutination zone 301 to the outer edge of the test readout zones 302, 303, 304, 205 is approximately 0.5 cm.
  • the ⁇ 300 was optimized to operate on approximately 7 ⁇ , whole blood samples, which corresponds to the amount of blood one could easily obtain with a finger prick and to the volume of blood sample required for many rapid diagnostic tests currently available in resource-limited settings.
  • test readout zones 302, 303, 304, 305 of the ⁇ 300 were made in a rectangular shape to simplify analysis of the color change in the test readout zones 302, 303, 304, 305.
  • the rectangular shape of the test readout zones 302, 303, 304, 305 of the ⁇ 300 design enables their automated selection when color change quantification is done by scanning and computer analysis.
  • the test readout zones may be of any shape.
  • the ⁇ 8 300 may be fabricated by printing the pattern of many ⁇ 8 300 (for example, arranged in an array) onto chromatography paper (for example, Whatman No. 1 chromatography paper, Piscataway, NJ) using a solid-ink (wax) printer (for example, a Phaser 8560N, Xerox, Norwalk, CT) and then heating the patterned paper on a hot plate at 150 °C for 3 minutes, and allowing said paper to cool to room temperature to enable the formation of hydrophobic barriers through the full thickness of the paper.
  • chromatography paper for example, Whatman No. 1 chromatography paper, Piscataway, NJ
  • a solid-ink (wax) printer for example, a Phaser 8560N, Xerox, Norwalk, CT
  • the melting process results in widening of the printed line, which was accounted for when originally designing the pattern of the ⁇ .
  • the ⁇ 300 is then functionalized by spotting (i) a solution of anti-A,B antibodies onto the agglutination zone 301, preferably of a volume in the range of 1-20 (ii) reagents of the colorimetric assay 301, preferably of a volume in the range of 1-20 L, onto each of the three of the test readout zones 302, 303, 304, and (iii) phosphate buffered saline 301 , preferably of a volume in the range of 1 - 20 ⁇ , onto one remaining test readout zone 305.
  • the test readout zone 305 treated with phosphate buffered saline is used for color change calibration.
  • Each functionalized ⁇ 300 is then allowed to dry before further use.
  • test readout zones 404, 405, 406, 407 where the analyte of interest of the plasma reacts with the reagents of the colorimetric assay producing a color change proportional to the concentration of the analyte of interest of the plasma.
  • test readout zones 404, 405, 406 are functionalized with the reagent of the colorimetric assay, thus resulting in a color change in those readout zones 404, 405, 406, and test readout zone 407 is not functionalized with the reagent of the colorimetric assay (and may instead be functionalized with phosphate buffered saline), thus acting as a control, and not resulting in a color change.
  • Example 1 Determination of Plasma Glucose Concentration in Whole Blood Samples
  • a ⁇ with RBC agglutination-based plasma separation was tested using an assay for plasma glucose as an example.
  • glucose oxidase catalyzes oxidation of glucose present in the sample of plasma to yield hydrogen peroxide (H2O2).
  • Horseradish peroxidase then catalyzes the reaction of 3 ⁇ 4(3 ⁇ 4 with potassium iodide, which results in brown color.
  • the intensity of the color change is proportional to the amount of H 2 0 2 produced, and thus to the amount of glucose.
  • FIG. 5 shows the calibration curve for the dependence of the color change on the concentration of glucose in plasma within the physiological range (50 - 200 mg/dL).
  • the three test readout zones 404, 405, 406 were each functionalized with 1 ⁇ of a solution consisting of potassium iodide (0.6M in deionized water), starch (0.3g/mL in saturated salt solution), glucose oxidase (100 U/mL in 0.1M potassium phosphate, pH 7.4, 0.05 M NaCl, 5 mM cholic acid, 0.1% Triton® X- 100), and horseradish peroxidase (20 U/mL in 0.1M potassium phosphate, pH 7.4, 0.05 M NaCl, 5 mM cholic acid, 0.1% Triton® X- 100).
  • the fourth test readout zone 407 was treated with 1 ⁇ , of phosphate buffered saline to control for changes in brightness and background color.
  • the agglutination zone 402 was functionalized with 7 of Seraclone Anti- ⁇ , ⁇ (AB03) clones BS 63/BS 85 (Biotest Medical Diagnostics GmbH, Germany). All reagents were allowed to dry before use of the ⁇ 400.
  • the color change value was converted into the plasma glucose concentration using the calibration curve for the assay (as shown in FIG. 5).
  • a smart phone equipped with a digital camera could also be used to complete this part of the assay.
  • the concentration of glucose in the whole blood sample was 89.5 mg/dL when measured with the ⁇ ) 400, and 82.5 mg/dL when measured independently using a conventional spectrophotometer (NanoDrop 1000).
  • This experiment used anti- ⁇ , ⁇ antibodies to induce RBC agglutination in whole blood samples obtained from volunteers with blood type A, B or AB.
  • blood type O approximately 44% of human population overall
  • Antigen H is present on the surface of all RBCs, including those with blood type 0 except those of Oh "Bombay phenotype" (less than 0.0004% of human population).
  • Antigen H is the precursor of antigen A and antigen B, and depending on the person's ABO blood type, it is converted into either antigen A or antigen B, or both.
  • RBCs of type A, B or AB have significantly less of antigen H than RBCs of type O, and we speculate that anti-H IgM antibodies would induce strong agglutination of type O RBCs and weak agglutination of type A, B or AB RBCs.
  • anti-H IgM antibodies would induce strong agglutination of type O RBCs and weak agglutination of type A, B or AB RBCs.
  • IgM antibodies reactive to antigens A, B and H either as a mixture of anti-H and anti- ⁇ , ⁇ or a single anti-ABH antibody
  • Another aspect of the present invention provides a diagnostic device and method for separating Hb A, C and F from deoxy-Hb S in small samples of whole blood contained entirely within a ⁇ in order to detect the presence of sickling hemoglobin in a blood sample.
  • Hb solubility assays such as SickleDex (SickleDexTM, Streck, Omaha, NE), that use saponin to chemically lyse RBCs in the blood sample, releasing Hb into solution where, in the presence of sodium hydrosulfite (an inexpensive and safe reducing agent), the freed Hb is converted to deoxy-Hb.
  • deoxy-Hb S changes conformation, polymerizes and precipitates, visibly clouding the solution (the solubility of non-sickling forms of Hb remains unaffected). Because of the polymerization, Hb S molecules agglutinate to form large supra-molecular agglomerates, which significantly increases their effective size with respect to the other types of Hb.
  • Hb solubility assays such as the SickleDex
  • SCT SCT
  • Hb SS SCD patients
  • SC SCD patients
  • One aspect of the present invention is a ⁇ addressing the aforementioned problem by using agglutination to separate Hb S from Hb A, C and F.
  • a drop of whole blood mixed with the components of a Hb solubility assay deposited onto a substrate will result in polymerized deoxy-Hb S (resulting from the release of Hb into solution where, in the presence of sodium hydrosulfite, the freed Hb is converted to deoxy- Hb and polymerizes).
  • the substrate may be paper, specifically chromatography paper, cloth, string or any other material with wicking or capillary properties.
  • FIG. 6 illustrates the operating principle of the paper-based Hb solubility assay schematically.
  • a drop of blood between 10-50 ⁇ , in volume, is added to an agglutinating agent, in this instance, SickleDex solution.
  • the SickleDex solution is added so that the volume ratio for the blood sample to SickleDex solution is 1 :20.
  • a droplet of this mixture 601 is deposited onto the paper substrate 602 of the ⁇ 600.
  • the overall diameter of the blood stain and the diameter of its center red spot 603 are determined by the volume of the droplet 601 deposited onto the paper substrate 602, and are independent of the type of the sample.
  • the color intensity of the pink ring 604 strongly correlates with the concentration of soluble forms of Hb (e.g. Hb A, F or C) present in the blood sample.
  • FIG. 7A illustrates the design of a ⁇ utilizing agglutination to separate Hb S from whole blood.
  • the ⁇ 700 includes square shaped hydrophobic barriers 702 and an agglutination zone 701 in the center region.
  • the square pattern of the hydrophobic barriers 702 of the ⁇ 700 was designed to limit the spread of blood from one ⁇ 700 to another, thus preventing the potential cross-contamination of samples.
  • the 45° alignment lines 703 in each corner of the ⁇ 700 provided the operator with visual guides for depositing the sample droplets in the agglutination zone 701 in the center of the ⁇ 700.
  • the simplistic design of the ⁇ 700 also significantly simplifies automated image analysis used to digitize and analyze the blood stain pattern.
  • the pattern of the hydrophobic barriers 702 were drawn in black lines on white background using illustration software (for example, Canvas 11, ACD Systems International Inc., Seattle, WA), and then printed on sheets of chromatography paper (for example, No. 1, Whatman, Piscataway, NJ) using a solid- ink printer (for example, a Phaser 8560N, Xerox, Norwalk, CT).
  • the printed chromatography paper was heated on a hot plate (150°C, 3 minutes) above the melting point of the wax to enable the formation of hydrophobic barriers 702 through the full thickness of the paper. The melting process resulted in widening of the printed line, which was accounted for when originally designing the pattern of the ⁇ 700.
  • a small volume approximately 10-50 ⁇ of whole blood, is gently mixed with the SickleDex solution at a 1 :20 ratio by volume, 5 minutes allowed to elapse, and then a 20 droplet of the mixture is deposited onto the center of the ⁇ 600.
  • the droplet spreads radially from the center through the paper substrate 602, forming a characteristic blood stain pattern.
  • the resulting blood stain is then digitized with a portable scanner and analyzed.
  • the blood stain pattern analyzed using an image processing algorithm.
  • the quantification of the blood stain is significantly simplified by the natural symmetry of the blood stain.
  • the computer algorithm automatically detects the geometric center of the stain, and the image is rotated with a 1° step about the center to collect 360 independent one-pixel-wide horizontal line scans of the blood stain (one such line scan is illustrated by the dashed line 605). These line scans are then averaged to obtain a single curve representative of the pattern of the red color intensity change from the center of the blood stain to its periphery. Examples of such curves for blood samples containing Hb AA, Hb AS and Hb SS are shown in FIG. 7B.
  • the red color intensity curves have approximately the same overall profile for all types of blood samples.
  • the color intensity increases gradually from the center of the stain, reaching maximum at the interface of the center spot and the peripheral pink ring.
  • This characteristic change in color across the center spot is speculated to be due to the transport of polymerized Hb S agglomerates with the radial outflow of liquid towards the contour left by the deposition of the original droplet.
  • the color of the pink ring is relatively uniform, fading into the background at the outer edge of the stain.
  • the uniformity in color of the pink ring is speculated to be due to the fact that the color of the pink ring is determined by the concentration of the soluble forms of Hb, which remain uniformly dissolved at the molecular level in the high-phosphate buffer solution.
  • Example 2 Classification of Blood Samples as Healthy, SCT or SCD
  • the ⁇ of the present invention was used on one normal (Hb AA), one SCT (Hb AS) and one SCD (Hb SS) blood sample as representative examples.
  • Hb AA normal human venous blood
  • SCD SCT
  • Hb AS SCT
  • Hb AS SCD
  • the Hb A, F, C and S content of SCD samples was determined via hemoglobin electrophoresis as a part of standard patient care.
  • SCT blood samples were collected from biological parents (usually mothers) of SCD patients. SCT samples with hematocrit values lower than 25% (indicating anemia) were excluded.
  • the SickleDex solution (SickleDexTM, Streck, Omaha, NE) used in this experiment is a commercially available test kit that consists of two components: (i) saponin and sodium hydrosulfite supplied as dry reagent power, and (ii) 2.3M potassium phosphate solubility buffer with 0.1% 2-chloroacetamide.
  • the contents of one vial containing the reagent powder were added to one bottle of the solubility buffer (as provided by the manufacturer) and dissolved completely with vigorous agitation.
  • the solution of the Hb solubility assay was mixed with blood at 1 :20 ratio by volume.
  • the scanned images were analyzed with an image algorithm (MATLAB ® , The Math Works Inc., Natick, MA) and digitized to produce the red color intensity profiles shown in FIG. 7B. It took about 10 minutes to complete all operations of the assay, including the introduction of the sample onto the ⁇ , the formation of the blood stain, scanning of the images and finally the automated image analysis. In contrast, a standard Hb electrophoresis test normally used to diagnose SCD takes at least 2 hours and often as long as about a week. [0074] Referring still to FIG.
  • the center spot 704 of the blood stain produced by SCT blood was significantly darker and the pink ring 705 on the periphery was significantly lighter than that of the normal sample.
  • the center spot 704 of the blood stain produced by SCD blood was the darkest of the three samples, and the pink ring 705 on the periphery of the ⁇ was barely visible.
  • Hb S which polymerizes when deoxygenated in a concentrated phosphate buffer
  • Hb AS SCT subjects
  • Hb SS SCD subjects
  • the SCD (Hb SS) sample which had the highest fraction of Hb S and the lowest fraction of soluble Hb (e.g. Hb A, F or C), produced the darkest center spot and a practically invisible pink ring on the periphery.
  • FIG. 8A shows the normalized color intensity profiles of blood stains for samples from normal (Hb AA), SCT (Hb AS) and SCD (Hb SS, Hb Sp, Hb SC) subjects.
  • the color intensity profiles were normalized by the total area under the curve (which reflects the Hb concentration in the original sample) to account for the differences in hematocrit among subj ects, and then averaged over all subjects within each Hb genotype.
  • the normalized color intensity at a distance of 5 mm from the center of the blood stain (dubbed SCD index and shown by the dashed line in FIG.
  • the physical meaning of the SCD index is the fraction of Hb in the sample that remains soluble when deoxygenated in concentrated phosphate buffer.
  • FIG. 8B we used the SCD index as a quantitative metric to differentiate samples among Hb genotypes.
  • the dots represent individual blood samples for each blood genotype (Hb AA (3), Hb AS (3), Hb SS (6), Hb S (1), and Hb SC(4)) and the SCD index value for that individual sample.
  • each blood genotypes Hb AA, Hb AS, Hb SS and Hb SC correspond to the mean of the samples (the center horizontal line) and one standard deviation above and below the mean (the top and bottom horizontal lines, respectively).
  • Our experiment only included one sample for blood genotype Hb Sfi, and as such, that sample includes only a mean and no standard deviation could be calculated.
  • FIG. 8B it can be observed that the majority of the samples analyzed using this method resulted in SCT values within one deviation from the mean. From FIG. 8B, one can also see that the SCD index for normal (Hb AA) samples was significantly higher (p ⁇ 0.001) than for any other type of samples tested (either SCT or SCD).
  • the SCD index for blood samples from SCT (Hb AS) individuals was significantly higher (p ⁇ 0.001) than for patients with Hb SS and Hb Sp° (the two most common and severe forms of SCD), and distinctively (although less significantly) higher (p ⁇ 0.05) than that for patients with Hb SC (a less common, milder form of SCD).
  • Hb SS / S group and Hb SC was also highly significant (p ⁇ 0.001 ), positioning the patients with Hb SC between generally healthy SCT individuals and patients with more severe forms of SCD in terms of the SCD index.
  • this SCT index value proves effective for identifying a blood sample as being normal (Hb AA), SCT (Hb AS) and SCD (Hb SS, Hb Sp, Hb SC).
  • the ⁇ of the instant invention may be used to diagnose the following diseases and infections using the following corresponding agglutinating agents: Acquired myasthenia gravis and Acetylcholine Receptor Antibody; Mycoplasma pneumoniae and cold agglutinins; Infectious mononucleosis and cold agglutinins; Influenza and cold agglutinins; Nonbacterial infection and cold agglutinins; Collagen vascular diseases and cold agglutinins; Cirrhosis and cold agglutinins; Leukemia, lymphoma, and multiple myeloma and cold agglutinins; Salmonella and febrile agglutinins; Rickettsia and febrile agglutinins; Brucellosis and febrile agglutinins; Tularemia and febrile agglutinins; Leukemia and febrile agglutinins; Lymp

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biochemistry (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Diabetes (AREA)
  • Ecology (AREA)
  • Biophysics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne un dispositif utilisant l'agglutination et son procédé d'utilisation pour diagnostiquer des maladies ou des affections. Le dispositif de diagnostic peut comprendre un substrat présentant des pores, une zone d'agglutination et une zone de lecture de test, ladite zone d'agglutination étant fonctionnalisée par un agent d'agglutination pour provoquer l'agglutination de l'échantillon.
PCT/US2012/064856 2011-11-10 2012-11-13 Test diagnostique à base de papier WO2013071301A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
US14/357,074 US20140295472A1 (en) 2011-11-10 2012-11-13 Paper Based Diagnostic Test
KR1020147015765A KR101659529B1 (ko) 2011-11-10 2012-11-13 종이기반 진단 테스트
EP12848168.6A EP2776831A4 (fr) 2011-11-10 2012-11-13 Test diagnostique à base de papier
CA2855363A CA2855363C (fr) 2011-11-10 2012-11-13 Test diagnostique a base de papier
CN201280062780.1A CN104126120B (zh) 2011-11-10 2012-11-13 纸基诊断测试
US15/215,107 US10908152B2 (en) 2011-11-10 2016-07-20 Paper based diagnostic test

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201161558009P 2011-11-10 2011-11-10
US61/558,009 2011-11-10
US201261692994P 2012-08-24 2012-08-24
US61/692,994 2012-08-24

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US14/357,074 A-371-Of-International US20140295472A1 (en) 2011-11-10 2012-11-13 Paper Based Diagnostic Test
US15/215,107 Continuation US10908152B2 (en) 2011-11-10 2016-07-20 Paper based diagnostic test

Publications (1)

Publication Number Publication Date
WO2013071301A1 true WO2013071301A1 (fr) 2013-05-16

Family

ID=48290684

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2012/064856 WO2013071301A1 (fr) 2011-11-10 2012-11-13 Test diagnostique à base de papier

Country Status (6)

Country Link
US (2) US20140295472A1 (fr)
EP (1) EP2776831A4 (fr)
KR (1) KR101659529B1 (fr)
CN (1) CN104126120B (fr)
CA (1) CA2855363C (fr)
WO (1) WO2013071301A1 (fr)

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101493051B1 (ko) 2014-03-07 2015-02-16 충남대학교산학협력단 3차원 종이기반 미세유체장비의 제작방법
WO2016019142A1 (fr) * 2014-07-30 2016-02-04 Case Western Reserve University Biopuces pour diagnostiquer des troubles de l'hémoglobine et surveiller les cellules sanguines
WO2016064545A1 (fr) * 2014-10-23 2016-04-28 Biomedomics, Inc. Procédés et dispositifs de dosage immunologique à écoulement latéral pour la détection simultanée de l'hémoglobine s, de l'hémoglobine c, et de l'hémoglobine a chez les nouveau-nés, les nourrissons, les enfants et les adultes
EP3115770A1 (fr) * 2015-07-07 2017-01-11 Commissariat à l'énergie atomique et aux énergies alternatives Procédé d estimation d'une quantité d analyte dans un liquide
US9597684B2 (en) 2015-01-16 2017-03-21 Xerox Corporation Method for making hydrophobic barriers in paper
US9782771B2 (en) 2015-01-16 2017-10-10 Xerox Corporation Method for making hydrophobic barriers requiring UV-light treatment
US20170336387A1 (en) * 2014-11-07 2017-11-23 The Johns Hopkins University Aggregation-assisted separation of plasma from whole blood
WO2018107129A1 (fr) 2016-12-09 2018-06-14 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr
WO2018170340A1 (fr) 2017-03-15 2018-09-20 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr pour la détection de virus
US10349589B2 (en) 2016-09-08 2019-07-16 Hemex Health, Inc. Diagnostics systems and methods
WO2019148206A1 (fr) 2018-01-29 2019-08-01 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr
WO2020072816A1 (fr) 2018-10-03 2020-04-09 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr pour la détection de fièvres hémorragiques
WO2020077135A1 (fr) 2018-10-10 2020-04-16 Dana-Farber Cancer Institute, Inc. Modulation de la résistance à des inhibiteurs de bcl -2
US10768166B2 (en) 2016-09-08 2020-09-08 Hemex Health, Inc. Diagnostics systems and methods
US11021740B2 (en) 2017-03-15 2021-06-01 The Broad Institute, Inc. Devices for CRISPR effector system based diagnostics
US11104937B2 (en) 2017-03-15 2021-08-31 The Broad Institute, Inc. CRISPR effector system based diagnostics
US11174515B2 (en) 2017-03-15 2021-11-16 The Broad Institute, Inc. CRISPR effector system based diagnostics
US11618928B2 (en) 2017-04-12 2023-04-04 The Broad Institute, Inc. CRISPR effector system based diagnostics for malaria detection
US11740203B2 (en) 2019-06-25 2023-08-29 Hemex Health, Inc. Diagnostics systems and methods
US11898142B2 (en) 2017-09-09 2024-02-13 The Broad Institute, Inc. Multi-effector CRISPR based diagnostic systems

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9686540B2 (en) * 2014-06-23 2017-06-20 Xerox Corporation Robust colorimetric processing method for paper based sensors
CN104730258B (zh) * 2015-03-12 2017-06-09 罗阳 一种便携式血型系统检测试纸及其检测方法
WO2017123668A1 (fr) 2016-01-12 2017-07-20 Trustees Of Tufts College Séparation de cellules sur la base de la taille et de l'affinité au moyen d'un dispositif microfluidique en papier
WO2019027963A1 (fr) * 2017-07-31 2019-02-07 Essenlix Corporation Dosages à interférence réduite
US20210263027A1 (en) * 2018-06-22 2021-08-26 Trustees Of Tufts College Patterned dried blood spot cards and related articles and methods
BR102019004230A2 (pt) * 2019-02-28 2020-09-08 Univ Do Vale Do Rio Dos Sinos Unisinos dispositivo analítico, processo de obtenção e uso do mesmo
WO2021019553A1 (fr) * 2019-07-27 2021-02-04 Indian Institute Of Technology, Kharagpur Système d'estimation d'hémoglobine sanguine basé sur un téléphone intelligent
US20220020481A1 (en) 2020-07-20 2022-01-20 Abbott Laboratories Digital pass verification systems and methods
US11498069B2 (en) 2020-08-31 2022-11-15 International Business Machines Corporation Rapid testing device for wastewater analysis
CN113903245A (zh) * 2021-10-08 2022-01-07 西安理工大学 一种比色型食品新鲜度智能指示标签及其制备方法
WO2023230305A1 (fr) * 2022-05-27 2023-11-30 Regents Of The University Of Minnesota Systèmes et procédés de dépistage de population pour la détection précoce de maladies chroniques

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090039130A (ko) * 2007-10-17 2009-04-22 한국전자통신연구원 랩온어칩 및 그 구동 방법
US20100140110A1 (en) * 2008-12-05 2010-06-10 Nanoivd, Inc. Microfluidic-based lab-on-a-test card for a point-of-care analyzer
WO2011000047A1 (fr) * 2009-06-30 2011-01-06 Monash University Analyse chimique quantitative et à étalonnage automatique utilisant des systèmes microfluidiques à base de papier
EP2281631A1 (fr) * 2009-08-07 2011-02-09 Atonomics A/S Système modulaire de préparation d'échantillons microfluidiques et procédé de mélange et de fourniture d'un échantillon fluide
KR20110091106A (ko) * 2010-02-05 2011-08-11 엘지전자 주식회사 혈액 샘플 분석 카트리지 및 카트리지 리더기
WO2011112023A2 (fr) * 2010-03-12 2011-09-15 주식회사 나노엔텍 Puce pour la séparation de cellules sanguines

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2031975A1 (fr) * 1990-01-12 1991-07-13 Brian R. Barkes Appareil et methode de separation et d'analyse du sang entier
US6168956B1 (en) * 1991-05-29 2001-01-02 Beckman Coulter, Inc. Multiple component chromatographic assay device
EP0535485B1 (fr) * 1991-10-03 1997-07-16 Bayer Corporation Dispositif et procédé pour la séparation et l'analyse du sang entier
US5660798A (en) * 1993-04-20 1997-08-26 Actimed Laboratories, Inc. Apparatus for red blood cell separation
US5753452A (en) * 1996-04-04 1998-05-19 Lifescan, Inc. Reagent test strip for blood glucose determination
CN1950700A (zh) * 2004-04-30 2007-04-18 爱科来株式会社 检体分析工具
KR100753595B1 (ko) * 2005-07-08 2007-08-30 세호로보트산업 주식회사 휴대단말기를 이용한 임상화학 측정 시스템 및 그 방법
CN101578520B (zh) * 2006-10-18 2015-09-16 哈佛学院院长等 基于形成图案的多孔介质的横向流动和穿过生物测定装置、及其制备方法和使用方法
CN100549678C (zh) * 2007-04-20 2009-10-14 上海农业信息有限公司 一种用农药残留检测仪进行农药残留检测的方法
SE531873C2 (sv) * 2007-11-12 2009-09-01 Lifeassays Ab Anordning för biokemisk bearbetning och analys av provvätska
AU2009228091B2 (en) * 2008-03-27 2015-05-21 President And Fellows Of Harvard College Three-dimensional microfluidic devices
CN101614736A (zh) * 2008-06-27 2009-12-30 北京华大吉比爱生物技术有限公司 应用新型全血样品垫处理方法的免疫层析检测试剂盒
WO2010102279A1 (fr) * 2009-03-06 2010-09-10 President And Fellows Of Harvard College Dispositifs électrochimiques microfluidiques
US20120322086A1 (en) * 2009-09-24 2012-12-20 Monash University Testing device for identifying antigens and antibodies in biofluids
CN102821861B (zh) * 2010-02-03 2015-03-25 哈佛大学校长及研究员协会 用于多重测定的装置和方法
CN201697916U (zh) * 2010-06-10 2011-01-05 桂林中辉科技发展有限公司 自动提示比色时间的目测尿糖试纸条
CN201935921U (zh) * 2011-01-30 2011-08-17 南京基蛋生物科技有限公司 一种带有u型过滤结构及三层滤血膜的胶体金全血检测卡
CN102182116B (zh) * 2011-03-14 2012-12-12 东莞南方医大代谢医学研发有限公司 一种收集生物样本的滤纸制备及运用方法
MX354260B (es) * 2011-07-22 2018-02-21 Access Bio Inc Una tira de almohadilla individual para un ensayo de flujo lateral mejorado y un dispositivo de análisis que usa la misma.

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090039130A (ko) * 2007-10-17 2009-04-22 한국전자통신연구원 랩온어칩 및 그 구동 방법
US20100140110A1 (en) * 2008-12-05 2010-06-10 Nanoivd, Inc. Microfluidic-based lab-on-a-test card for a point-of-care analyzer
WO2011000047A1 (fr) * 2009-06-30 2011-01-06 Monash University Analyse chimique quantitative et à étalonnage automatique utilisant des systèmes microfluidiques à base de papier
EP2281631A1 (fr) * 2009-08-07 2011-02-09 Atonomics A/S Système modulaire de préparation d'échantillons microfluidiques et procédé de mélange et de fourniture d'un échantillon fluide
KR20110091106A (ko) * 2010-02-05 2011-08-11 엘지전자 주식회사 혈액 샘플 분석 카트리지 및 카트리지 리더기
WO2011112023A2 (fr) * 2010-03-12 2011-09-15 주식회사 나노엔텍 Puce pour la séparation de cellules sanguines

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CARRILHO, E. ET AL.: "Understanding Wax Printing: A Simple Micropatterning Pr ocess for Paper-Based Microfluidics", ANALYTICAL CHEMISTRY, vol. 81, no. 16, 15 July 2009 (2009-07-15), pages 7091 - 5, XP002585317 *
See also references of EP2776831A4 *
YANG, X. ET AL.: "Integrated separation of blood plasma from whole blood for microfluidic paper-based analytical devices", LAB ON A CHIP, vol. 12, 17 November 2011 (2011-11-17), pages 274 - 280, XP055149746 *

Cited By (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101493051B1 (ko) 2014-03-07 2015-02-16 충남대학교산학협력단 3차원 종이기반 미세유체장비의 제작방법
WO2016019142A1 (fr) * 2014-07-30 2016-02-04 Case Western Reserve University Biopuces pour diagnostiquer des troubles de l'hémoglobine et surveiller les cellules sanguines
WO2016064545A1 (fr) * 2014-10-23 2016-04-28 Biomedomics, Inc. Procédés et dispositifs de dosage immunologique à écoulement latéral pour la détection simultanée de l'hémoglobine s, de l'hémoglobine c, et de l'hémoglobine a chez les nouveau-nés, les nourrissons, les enfants et les adultes
US20170336387A1 (en) * 2014-11-07 2017-11-23 The Johns Hopkins University Aggregation-assisted separation of plasma from whole blood
US10788480B2 (en) * 2014-11-07 2020-09-29 The Johns Hopkins University Aggregation-assisted separation of plasma from whole blood
US9597684B2 (en) 2015-01-16 2017-03-21 Xerox Corporation Method for making hydrophobic barriers in paper
US9782771B2 (en) 2015-01-16 2017-10-10 Xerox Corporation Method for making hydrophobic barriers requiring UV-light treatment
US10346983B2 (en) 2015-07-07 2019-07-09 Commissariat A L'energie Atomique Et Au Energies Alternatives Method for estimating an amount of analyte in a fluid
EP3115770A1 (fr) * 2015-07-07 2017-01-11 Commissariat à l'énergie atomique et aux énergies alternatives Procédé d estimation d'une quantité d analyte dans un liquide
FR3038723A1 (fr) * 2015-07-07 2017-01-13 Commissariat Energie Atomique Procede d’estimation d’une quantite d’analyte dans un liquide
US10049453B2 (en) 2015-07-07 2018-08-14 Commissariat A L'energie Atomique Et Aux Energies Alternatives Method for estimating an amount of analyte in a fluid
US10375909B2 (en) 2016-09-08 2019-08-13 Hemex Health, Inc. Diagnostics systems and methods
US11701039B2 (en) 2016-09-08 2023-07-18 Hemex Health, Inc. Diagnostics systems and methods
US11656220B2 (en) 2016-09-08 2023-05-23 Hemex Health, Inc. Diagnostics systems and methods
US10349589B2 (en) 2016-09-08 2019-07-16 Hemex Health, Inc. Diagnostics systems and methods
US10768166B2 (en) 2016-09-08 2020-09-08 Hemex Health, Inc. Diagnostics systems and methods
WO2018107129A1 (fr) 2016-12-09 2018-06-14 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr
EP4119663A1 (fr) 2016-12-09 2023-01-18 The Broad Institute, Inc. Diagnostic basé sur un système effecteur de crispr
US11104937B2 (en) 2017-03-15 2021-08-31 The Broad Institute, Inc. CRISPR effector system based diagnostics
US11021740B2 (en) 2017-03-15 2021-06-01 The Broad Institute, Inc. Devices for CRISPR effector system based diagnostics
US11174515B2 (en) 2017-03-15 2021-11-16 The Broad Institute, Inc. CRISPR effector system based diagnostics
WO2018170340A1 (fr) 2017-03-15 2018-09-20 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr pour la détection de virus
US11618928B2 (en) 2017-04-12 2023-04-04 The Broad Institute, Inc. CRISPR effector system based diagnostics for malaria detection
US11898142B2 (en) 2017-09-09 2024-02-13 The Broad Institute, Inc. Multi-effector CRISPR based diagnostic systems
WO2019148206A1 (fr) 2018-01-29 2019-08-01 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr
WO2020072816A1 (fr) 2018-10-03 2020-04-09 The Broad Institute, Inc. Diagnostics basés sur un système effecteur crispr pour la détection de fièvres hémorragiques
WO2020077135A1 (fr) 2018-10-10 2020-04-16 Dana-Farber Cancer Institute, Inc. Modulation de la résistance à des inhibiteurs de bcl -2
US11740203B2 (en) 2019-06-25 2023-08-29 Hemex Health, Inc. Diagnostics systems and methods

Also Published As

Publication number Publication date
CN104126120B (zh) 2017-12-08
EP2776831A4 (fr) 2015-06-03
US20140295472A1 (en) 2014-10-02
US20170023556A1 (en) 2017-01-26
CN104126120A (zh) 2014-10-29
US10908152B2 (en) 2021-02-02
KR20140109880A (ko) 2014-09-16
CA2855363A1 (fr) 2013-05-16
KR101659529B1 (ko) 2016-09-23
EP2776831A1 (fr) 2014-09-17
CA2855363C (fr) 2019-02-12

Similar Documents

Publication Publication Date Title
US10908152B2 (en) Paper based diagnostic test
EP2780705B1 (fr) Procédés et systèmes de détection d'un analyte dans un échantillon
RU2503009C2 (ru) Устройство и способ для разделения и анализа крови
US8377398B2 (en) Methods and compositions related to determination and use of white blood cell counts
Vyas et al. Development of immunochromatographic strip test using fluorescent, micellar silica nanosensors for rapid detection of B. abortus antibodies in milk samples
JP5335802B2 (ja) 潜血を検出する方法および装置
US9097712B2 (en) Flow-through cell counting assay
Ayele et al. Evaluation of circulating cathodic antigen (CCA) strip for diagnosis of urinary schistosomiasis in Hassoba school children, Afar, Ethiopia
JP2007139556A (ja) 新規な分析方法およびキット
CN106596968A (zh) 一种检测尿微量白蛋白斑点金渗滤试剂盒及其应用
US20210132049A1 (en) Simultaneous Parallel Signal Amplification and Analyte-Ligand Capture Functions
WO2017013267A1 (fr) Procédé pour évaluer les récepteurs de surface cellulaire des cellules sanguines
Bystryak et al. A flow-through cell counting assay for point-of-care enumeration of CD4 T-cells
WO1997023781A1 (fr) Procede et dispositif d'analyse de matieres fecales
WO2008104081A1 (fr) Dispositif de dosage immunologique parallèle
CN113281504B (zh) 一种新型免疫层析检测装置
EP1933148B1 (fr) Récipient pour la détection immunochromatographique de plusieurs paramètres urinaires
WO2023180899A1 (fr) Dispositif et procédé de détection de diarrhée
WO2020012323A2 (fr) Procédé d'identification d'une infection bactérienne des voies urinaires chez l'enfant de moins de 2 ans
CN118191308A (zh) 一种感染关节液的检测平台及检测方法
Wang et al. Emerging microscale technologies for global health: CD4 counts
CA3030069A1 (fr) Dispositif d'analyse et procede d'evaluation des cellules sanguines
Parija Changing Trends in the Diagnosis of Parasitic Diseases–An Indian Scenario
BR102013003143A2 (pt) PROCESSO DE FABRICAÇÃO DE DISPOSITIVOS ANALÍTICOS CONTENDO ARRANJOS MÚLTIPLOS DE MICROZONAS, OU MICROWELLS, PARA MONITORAMENTO, ENSAIOS E DIAGNÓSTICOS CLÍNICOS NO POINT-OF-CARE (PoC)

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12848168

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 14357074

Country of ref document: US

ENP Entry into the national phase

Ref document number: 2855363

Country of ref document: CA

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2012848168

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 20147015765

Country of ref document: KR

Kind code of ref document: A